TABLE 4.
Complementation between individually cloned tou genes and the tou gene clusters carrying the corresponding mutation
| Plasmid(s)a | tou genesb | IPTG addition | Toluene/o-xylene monooxygenase sp act (nmol min−1 mg of protein−1) on substrate
|
|||
|---|---|---|---|---|---|---|
| Step Ic
|
Step IId
|
|||||
| Toluene | o-Xylene | m-Cresol | 2,3-DMP | |||
| pMM3356 | ABCDEF | + | 10.00 | 4.71 | 6.25 | 3.50 |
| − | 2.30 | 0.00 | 2.30 | 2.00 | ||
| pMM3301 | ABCDEF | + | 0.00 | 0.00 | 0.34 | 0.00 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3302 | ABCDEF | + | 0.00 | 0.00 | 0.00 | 0.00 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3303 | ABCDEF | + | 0.72 | 0.24 | 1.28 | 1.12 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3304 | ABCDEF | + | 0.00 | 0.00 | 0.00 | 0.00 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3305 | ABCDEF | + | 0.00 | 0.00 | 0.00 | 0.00 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3306 | ABCDEF | + | 4.31 | 1.30 | 4.65 | 1.40 |
| − | 0.00 | 0.00 | 0.00 | 0.96 | ||
| pMM3301, pMZ1201 | ABCDEF + A | + | 2.23 | 1.23 | 1.57 | 2.47 |
| − | 0.00 | 0.00 | 0.00 | 0.70 | ||
| pMM3302, pMZ9002 | ABDEF + B | + | 4.16 | 0.75 | 2.61 | 0.83 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3303, pMZ1203 | ABCDEF + C | + | 2.94 | 0.78 | 5.42 | 2.08 |
| − | 0.00 | 0.00 | 0.00 | 0.00 | ||
| pMM3304, pMZ1204 | ABCDEF + D | + | 11.47 | 2.87 | 5.05 | 2.35 |
| − | 0.67 | 0.32 | 0.84 | 0.71 | ||
| pMM3305, pMZ1205 | ABCDEF + E | + | 5.52 | 1.83 | 6.94 | 2.79 |
| − | 0.89 | 0.26 | 0.77 | 0.69 | ||
| pMM3306, pMZ1006 | ABCDEF + F | + | 6.45 | 3.00 | 6.45 | 2.62 |
| − | 2.20 | 0.00 | 0.00 | 1.40 | ||
a
Construction of each plasmid is described in Materials and Methods. The host was E. coli DH5α.
b
The mutated genes are indicated in boldface.
c
Conversion of the specified hydrocarbon into a phenolic compound.
d
Consumption of the specified phenolic compound.